Adenoviral tumor diagnostics
Compositions and methods for detecting a cancer in a subject using a recombinant reporter adenovirus are described. In particular, recombinant adenovirus is used to diagnose a cancer in a patient and further used for screening compounds effective in treating the cancer in said patient.
1. A recombinant reporter adenovirus, comprising a first cancer cell reporter module, a second cancer cell reporter module and a cancer cell binding module, wherein said first cancer cell reporter module comprises a constitutive promoter active in tumor cells and non-tumor cells operably linked to a first reporter gene that expresses a first reporter gene phenotype in tumor cells and non-tumor cells, and said second cancer cell reporter module comprises a cancer responsive promoter operably linked to a second reporter gene that expresses a second reporter gene phenotype in tumor cells, and wherein said first reporter gene phenotype and said second reporter gene phenotype are detectably different.
2. The recombinant reporter adenovirus of claim 1 , further comprising an immune evasion module.
3. The recombinant reporter adenovirus of claim 1 , wherein said first reporter gene or said second reporter gene comprises a fluorescent reporter gene.
4. The recombinant reporter adenovirus of claim 1 , further comprising a third cancer cell reporter module, wherein said third cancer cell reporter module comprises a cancer responsive promoter operably linked to a third reporter gene that expresses a third reporter gene phenotype in tumor cells, wherein said first reporter gene phenotype, said second reporter gene phenotype, and said third reporter gene phenotype are each detectably different.
5. A method of detecting a cancer in a subject, said method comprising:
(i) administering the recombinant reporter adenovirus of claim 1 to a subject;
(ii) allowing said recombinant reporter adenovirus to infect a cancer cell within said subject thereby forming a reporter infected cancer cell;
(iii) obtaining from said subject a sample comprising said reporter infected cancer cell; and
(iv) detecting said reporter infected cancer cell thereby detecting a cancer in said subject.
6. The method of claim 5 , wherein said detecting comprises detecting a reporter gene phenotype.
7. A method of detecting a cancer in a subject, the method comprising:
(i) obtaining from a subject a sample comprising a cancer cell;
(ii) contacting the recombinant reporter adenovirus of claim 1 with said cancer cell;
(iii) allowing said recombinant reporter adenovirus to infect said cancer cell thereby forming a reporter infected cancer cell; and
(iv) detecting said reporter infected cancer cell thereby detecting a cancer in said subject.
8. The method of claim 7 , wherein said detecting comprises detecting a reporter gene phenotype.
9. A method of determining whether a test compound inhibits growth of a cancer cell from a cancer patient, said method comprising:
(i) obtaining from a subject a sample comprising a cancer cell;
(ii) contacting the recombinant reporter adenovirus of claim 1 with said cancer cell;
(iii) allowing said recombinant reporter adenovirus to infect said cancer cell thereby forming a reporter infected cancer cell;
(iv) allowing said reporter infected cancer cell sufficient time to grow;
(v) determining a level of growth of said reporter infected cancer cell; and
(vi) comparing said level to a control level, wherein a low level compared to said control level indicates said test compound inhibits growth of said cancer cell from said patient.
10. A kit for detecting cancer, for screening a cancer drug and/or for isolating a cancer cell, said kit comprising the recombinant reporter adenovirus of claim 1 and (i) reagents for separating cells from a tissue or cell sample from a subject; (ii) a cancer inhibiting compound; and/or (iii) a device for detecting an expressed reporter gene phenotype.
11. The recombinant reporter adenovirus of claim 1 , wherein the cancer cell binding module comprises an adenovirus fiber protein that binds CD46.
12. The recombinant reporter adenovirus of claim 1 , wherein the cancer cell binding module comprises an Ad3, Ad11 or Ad34 fiber knob.
13. The recombinant reporter adenovirus of claim 1 , wherein the cancer responsive promoter is active only in:
(a) pRb/p16 inactive cells;
(b) cells with an activated EGFR/RAS/RAF/MAPK pathway; or
(c) cells with active transforming growth factor (TGF)-β signaling.
14. The recombinant reporter adenovirus of claim 13 , wherein the cancer responsive promoter is E2F, serum response element (SRE) or a SMAD-responsive promoter.
15. The recombinant reporter adenovirus of claim 1 , wherein the first reporter gene and/or the second reporter gene encodes a fusion protein comprising a fluorescent protein and a protein that directs localization of the fusion protein to either the cytoplasm or the nucleus.
16. The recombinant reporter adenovirus of claim 15 , wherein the protein that directs location of the fusion protein comprises FOXO3 or CRTC2.
17. The recombinant reporter adenovirus of claim 1 , wherein the first reporter gene or the second reporter gene encodes a green fluorescent protein, a yellow fluorescent protein or a red fluorescent protein.
18. The recombinant reporter adenovirus of claim 17 , wherein the fluorescent protein comprises mCherry or tdTomato.